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Publication numberUS4576702 A
Publication typeGrant
Application numberUS 06/668,562
Publication dateMar 18, 1986
Filing dateNov 5, 1984
Priority dateNov 5, 1984
Fee statusLapsed
Also published asDE3539136A1
Publication number06668562, 668562, US 4576702 A, US 4576702A, US-A-4576702, US4576702 A, US4576702A
InventorsLawrence J. Peck, John H. Kreisher
Original AssigneeInternational Biotechnologies, Inc.
Export CitationBiBTeX, EndNote, RefMan
External Links: USPTO, USPTO Assignment, Espacenet
Analytical electroelution device
US 4576702 A
Abstract
A simplified electroelution receptacle designed to receive and process a plurality of small biological samples simultaneously facilitating efficient recovery of biological particles in a highly concentrated manner with a minimum of undesirable liquid.
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Claims(12)
We claim:
1. An electroelution device for removing and collecting charged biological particles from a plurality of small gel slices or samples simultaneously comprising at least two fluid tight compartments separated by a bridge element, said bridge element being formed with a plurality of spaced recesses defining sample supports, said bridge being further formed with a plurality of conduits, one conduit being individual to each sample support and each conduit having a first outlet communicating with a mating sample support and a second outlet communicating with one of said compartments.
2. The device of claim 1 in which the first and second outlets are offset from one another.
3. The device of claim 2 in which the first outlets are coplanar and are offset vertically from the second outlets.
4. The device of claim 3 in which the first outlets are elevated relative to the disposition of the second outlets.
5. The device of claim 4 in which the conduits define a generally V-shaped configuration between outlets.
6. The device of claim 5 in which a portion of each conduit is offset vertically relative to the elevation of all outlets.
7. The device of claim 6 in which the conduits are generally V-shaped and the apex of the V is lower in elevation than the elevation of all outlets.
8. The device of claim 7 in which the bridge element is fitted with a valve means which includes ports which communicates selectively with both compartments and with the atmosphere.
9. The device of claim 8 in which the valve means is settable (a) to a first position in which opposed ports provide communication between both compartments, (b) to a second position in which said communication is cut-off, (c) to a third position in which said opposed ports and thus the compartments communicate with the atmosphere.
10. The device of claim 9 in which the bridge element is provided with an overflow port which provides a by-pass to the atmosphere independently of the valve means.
11. The device of claim 10 in which each compartment is fitted with an electrode leading to a fixed electrical connection mounted in the receptacle.
12. The device of claim 11 in which at least one compartment is fitted with cooling tubes for the circulation of coolant for controlling temperature.
Description
BACKGROUND OF THE INVENTION

The present invention relates to electrophoresis and relates in particular to a novel receptacle or apparatus for removing (eluting) and collecting charged particles of biological substances from gel slices in highly concentrated forms and with a bare minimum of undesirable liquid.

The procedure is referred to hereinafter as electroelution.

The present invention is an improvement over a copending application filed Nov. 5, 1984 by Peck et al. entitled Preparatory Electroelution Device bearing Ser. No. 668,571 both applications having a common Assignee.

The isolation of DNA, RNA, carbohydrates and proteins from electrophoresis gels has been a long standing problem. Several methods are available for recovering nucleic acids from acrylamide and agarose gels. These include (1) elution by diffusion, (2) extrusion by compression, (3) gel dissolution, and (4) electroelution. Most of these systems result in poor yield, degradation, contaminated end products, and are very cumbersome to operate. Electroelution is often the best method, although up to now, made difficult by the necessity for membranes, tube gels and related paraphernalia. In electroelution systems, the biologicals can stick to the membranes when dialysis tubing or membrane systems are used. A reverse current will partially release the material, but the process is not quantitative. Smaller fragments may pass through the membrane. Tube gels and related electrophoresis systems are cumbersome to operate.

The present invention provides an improved apparatus useful to carry out electroelution procedures in a manner which eliminates many of the difficulties encountered in prior art structures.

In the improved device an array of sample supports are provided facilitating the removal and collection of biological particles from a number of samples simultaneously.

Treatment of multiple samples simultaneously saves time of highly trained technical personnel and provides a series of results from a given family of samples in a very convenient fashion facilitating subsequent procedural steps.

Consequently it is a prime object of the present invention to provide an efficient electroelution device.

It is a further object to provide a device which is operative to recover and collect small particles or fragments of particles which would pass through and be lost in membrane arrangements of the prior art.

For example, the present invention is operable to concentrate masses of particles of the order of 10 to 20 micrograms in a liquid volume as small as 200 microliters.

A further feature of the invention is the provision of a novel electroelution receptacle structure which is accessable, convenient to operate, purge and keep clean.

SUMMARY OF THE INVENTION

An electroelution device or receptacle embracing certain principles of the invention may comprise at least two fluid tight compartments separated by a bridge element, said bridge element being formed with a plurality of spaced recesses defining sample supports, said bridge being further formed with a plurality of conduits, one conduit being individual to each sample support and each conduit having a first outlet communicating with a mating sample support and a second outlet communicating with one of said compartments.

BRIEF DESCRIPTION OF THE DRAWINGS

Other features and advantages of the present invention will become more apparent from an examination of the appended drawings, in which:

FIG. 1 is an exploded view in perspective of an electroelution device illustrating the principles of this invention;

FIG. 2 is a top plan view;

FIG. 3 is a front elevation of FIG. 2; and

FIG. 4 is an elevational view of the right side of FIG. 2.

DETAILED DESCRIPTION

Referring now in detail to the drawings the electroelution device, indicated generally by the reference numeral 11, includes a first compartment 12, a second compartment 13 separated by a bridge element 14.

The bridge element 14 is formed with an array of spaced recesses 16--16 defining sample supports or sample receptacles.

Each recess 16 is formed with a first cut-out or sluice way 17 directed to compartment 12 and a second, opposed cut-out 18 directed to an outlet 19 of a conduit 21.

The conduit 21 is generally V-shaped and terminates in an outlet 22 opening into compartment 13.

The outlets 19--19 lie generally in the same plane and are offset vertically above coplanar outlets 22--22.

The tip or low point of the conduit 21 is spaced vertically and below all outlets 19 and 22.

A three position valve, indicated generally at 23, is movable selectively from a first position in which compartments 12 and 13 communicate through the valve stem opening 20 and opposed bridge ports 24 and 26, to a second position in which the valve stem 27 cuts off compartment communication, to a third position in which both compartments 12 and 13 communicate to atmosphere through stem openings 28 and 19 and conduit 31 in the manner described in similar detail in said copending application.

The bridge element 14 also includes an overflow port 32 which by-passes valve 23 and also communicates with atmosphere through conduit 31.

Electrical connectors 33-34 supply current to mating electrodes 36 and 37 disposed in the bottom of each compartment.

A cover 38 hinged to rod 39 by U-shaped clips 41--41 is fitted with ferrules 42--42 providing access for the electrodes 36 and 37.

The cooperation between the cover 38 and head 43 of valve stem 25 and the control of the stroke of the stem 25 are as disclosed and described in said copending application.

In order to control temperature when temperature sensitive materials are involved, one or both of the compartments 12 and 13 are provided with coolant coils 44 indicated in dotted lines in FIG. 1.

A carpenter's level 45 with appropriate levelling screws 47--47 is provided to level the surface of fluid within compartments 12 and 13.

The electroelution device operates in the following fashion:

The valve 23 is elevated to its first position in which there is communication between both compartments 12 and 13, and the unit is levelled. Next, appropriate buffer liquid is introduced into one of the other compartments filling the compartments until overflow occurs to atmosphere at overflow port 32. This overflow occurrence establishes the proper level of buffer liquid.

Thereafter a specific salt of predetermined density (of substantially greater density than the density of the buffer liquid) is pipetted into each V-shaped conduit 21--21. This step displaces the lower gravity buffer liquid from the V-shaped conduits. Next samples are distributed into the array of sample supports 16--16.

The cover 38 is closed causing valve 23 to move to the second position cutting off communication between compartments 12 and 13.

Electrical potential is applied to connectors 33-34 and the electroelution procedures begins and continues for an appropriate interval.

Recovered particles collect upon the surface of the dense salt contained in the V-shaped conduits.

When the collection procedure is completed the power is disconnected and the valve 23 moved to its third position permitting buffer liquid to drain to atmosphere isolating the matter in the V-shaped conduits.

Thereafter the contents of the V-shaped conduits are removed by pipetting for further treatment.

As stated previously if the samples are heat sensitive it is sometimes necessary to run coolant through coolant tubes--to maintain desired temperature gradient.

It is to be understood that the invention is not limited to the illustrations described and shown herein, which are deemed to be merely illustrative of the best modes of carrying out the invention, and which are susceptible of modification of form, size, arrangement of parts and details of operation. The invention rather is intended to encompass all such modifications which are within its spirit and scope as defined by the claims.

Patent Citations
Cited PatentFiling datePublication dateApplicantTitle
US3989612 *Dec 26, 1974Nov 2, 1976The Upjohn CompanyBuffers, direct current
US4049534 *Apr 12, 1976Sep 20, 1977E-C Apparatus CorporationElectrophorectic apparatus for elution from preparative gels
SU552373A1 * Title not available
Non-Patent Citations
Reference
1Diekmann, S., "A Device to Flute Biomolecules Out of Gels", Electrophoresis '84, Editor: Volker Neuhoff, pp. 154-155, Jul. 1984.
2 *Diekmann, S., A Device to Flute Biomolecules Out of Gels , Electrophoresis 84, Editor: Volker Neuhoff, pp. 154 155, Jul. 1984.
Referenced by
Citing PatentFiling datePublication dateApplicantTitle
US4695650 *Sep 24, 1986Sep 22, 1987University Of Colorado FoundationLiquid crystal compounds and compositions containing same
US4725348 *May 3, 1985Feb 16, 1988Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften E.V.Electrophoretic transfer of biopolymers jmikokim permits simultaneous skimming and vacuuming; back pressure compensation
US4744918 *Jul 11, 1985May 17, 1988Merck Patent Gesellschaft Mit Beschrankter HaftungEsters,optical apparatus
US4747918 *Jun 18, 1986May 31, 1988William BellomyGel electrophoresis
US4773984 *Feb 2, 1987Sep 27, 1988Life Technologies, Inc.Ad-justable, self-locking supports; separation of dna or rna fragments
US4830725 *Aug 4, 1987May 16, 1989Life Technologies, Inc.Electrophoresis apparatus
US4859302 *Oct 22, 1987Aug 22, 1989President And Fellows Of Harvard CollegeElectroelution of macromolecules
US4959133 *Jan 23, 1989Sep 25, 1990Eastman Kodak CompanyField inversion electroblotting & electroelution
US5139637 *Mar 13, 1991Aug 18, 1992Macconnell William PPlasmid purification system and method
US5217593 *Jul 10, 1992Jun 8, 1993Macconnell William PNucleic acid purification system and method
US5405520 *Jan 27, 1994Apr 11, 1995Eastman Kodak CompanyConnectors for electrophoresis device
US5415758 *Nov 19, 1993May 16, 1995Theobald Smith Research Institute, Inc.Method and apparatus for electro-elution of biological molecules
US5779869 *Nov 22, 1996Jul 14, 1998Eastman Kodak CompanyLow fog electrophoresis device
US6063250 *May 15, 1998May 16, 2000C.C. ImexRunning tank assembly for electrophoresis
US6402915Jul 15, 1999Jun 11, 2002C.C. ImexElectrolytic cells; prevention tilting of lid
US6929730May 1, 2002Aug 16, 2005Cheng Sheng LeeTwo dimensional microfluidic gene scanner
US6974526May 1, 2002Dec 13, 2005Calibrant Biosystems, Inc.Simultaneously crushing four or more catalysis materials in four or more spatially discrete crushing zones of a parallel crusher, sieving each of the four or more catalysis materials; sieving the first-sieved particles
US7025864Mar 10, 2003Apr 11, 2006Elchrom Scientific A.G.Method and apparatus for recovering target molecules from a gel containing said target molecules
US7070682Jan 15, 2002Jul 4, 2006Cheng LeeMicrofluidic apparatus for performing gel protein extractions and methods for using the apparatus
US7074313Nov 22, 2002Jul 11, 2006Gene Bio-Application Ltd.Electroelution and/or dialysis using a particular gel
US7641780Mar 23, 2005Jan 5, 2010Calibrant Biosystems, Inc.Two-dimensional microfluidics for protein separations and gene analysis
WO1994001763A1 *Apr 19, 1993Jan 20, 1994William P MacconnellNucleic acid purification system and method
WO2001090731A2 *May 22, 2001Nov 29, 2001Gene Bio Applic LtdProcessing chamber with apertures for pipette access
Classifications
U.S. Classification204/613
International ClassificationB01D57/02, C12N13/00, C12M1/00, G01N27/447, G01N27/26
Cooperative ClassificationG01N27/44739
European ClassificationG01N27/447B3B
Legal Events
DateCodeEventDescription
May 26, 1998FPExpired due to failure to pay maintenance fee
Effective date: 19980318
Mar 15, 1998LAPSLapse for failure to pay maintenance fees
Feb 12, 1998REMIMaintenance fee reminder mailed
Jul 21, 1993FPAYFee payment
Year of fee payment: 8
Jul 27, 1989FPAYFee payment
Year of fee payment: 4
Nov 23, 1984ASAssignment
Owner name: INTERNATIONAL BIOTECHNOLOGIES, INC. 275 WINCHESTER
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNORS:PECK, LAWRENCE J.;KREISHER, JOHN H.;REEL/FRAME:004343/0158;SIGNING DATES FROM 19840914 TO 19841023